Diacetylenic glycerophosphatidylhydroxyethanol was synthesized from diacetylenic glycerophosphatidylcholine in the presence of phospholipase D from peanut seeds. The product structure was characterized by elemental analysis, IR, 1H NMR and 13C NMR. The effects of the reaction conditions on the enzyme-catalyzed reaction are discussed. The results show that the enzyme (activity) was greatly increased by adding SDS. The yield of the reaction was increased compared with that reported in the literature under the conditions of pH=5.6, temperature 30 ℃, Ca2+ 40mmol/L, (n(substrate )∶)n(SDS)=(2∶1), reaction time 8 h. The cost was lower. The melting point of the product was 69.5 ℃.